Table 1. Description of primer sequences, fluorescent labeling and length extension of PCR products amplified by modified primers.
locus name primer sequence (5´–3´) primer
concentration
in multiplex (μM) 		fluorescent
dye
(5’ F primer) 		extension of modified
amplicons
(bp)
source literature / GenBank accession number
multiplex A
EMPaS02 F: CTACTTCCATGATTGCCTCAC 0.2 FAM -
A / AY526619 R: AACATCCAGAACATCAACACAC      
*EMPaS10 long F: GCTAATATCAAATCCCAGCTCTC 0.2 VIC 127
A / AY526626 R: CTACTGGCTTGTGTTGTGTG      
EMPaS11 F: ACCACTTTGAGGAACTTGGG 0.2 FAM -
A / AY526627 R: CTGCCTGGAAGAGCAATAAC      
UCD-CH11 F: TGCTATTAGCTTAATGCCTCCC 0.3 PET -
C / NA R: ATGCTGATGTCATAAGGTGTGC      
UCD-CH31 F: TCCGCTTCTCTGTGAGTGTG 0.2 NED -
C / NA R: CGATAGTTTCCTTCCCAGACC  
*PS12A02 long F: GCCACCAATGGTTCTTCC 0.3 NED 7
E / AB476763 R: CAACCAAAGCACCAGATGC      
UDP98-412 F: AGGGAAAGTTTCTGCTGCAC 0.2 VIC -
B / AY074720 R: GCTGAAGACGACGATGATGA      
multiplex B  
EMPaS05 F: CATGTGCTTTCTCTGCCC 0.2 VIC -
A / AY526621 R: TCTTCTCAAGCAATTCCCC      
EMPaS06 F: AAGCGGAAAGCACAGGTAG 0.2 NED -
A / AY526622 R: TTGCTAGCATAGAAAAGAATTGTAG      
*EMPaS12 long F: TGTGCTAATGCCAAAAATACC 0.2 FAM 99
A / AY526628 R: GTGGAAGGCCATATTTTCGG      
*BPPCT 034 long F: CTACCTGAAATAAGCAGAGCCAT 0.3 PET 99
D / AF374945 R: GCTCGTAAGATTGCTGGTAGC      
multiplex C  
*UDP97-403 long F: CTGGCTTACAACTCGCAAGC 0.3 VIC 76
B / AY074723 R: CTCACAGACTCGACTCTTCACTTC      
*PceGA34 long F: GAACATGTGGTGTGCTGGTT 0.2 NED 100
E / NW007362363 R: CCCACTTTCATCAAAACAGTGAG      
*UDP98-411 long F: AAGCCATCCACTCAGCACTC 0.15 PET 108
B / AY074719 R: GCTGATGATGACGACGATGATG      
UCD-CH12 F: AGACAAAGGGATTCTGGGC 0.4 PET -
C / NA R: TTTCTGCCACAAACCTAATGG      
* – modified loci are tagged with suffix long, sequences of redefined reverse primers are highlighted in bold; NA – sequences not published in GeneBank database.
Source literature: A – Vaughan and Russel 2004, B –Testolin et al. 2000 (Schueller et al. 2006), C – Struss et al. 2003, D – Dirlewanger et al. 2002, E – Downey and Iezzoni 2000.
Table 2. Size ranges of PCR products in pseudo 15-plex.
fluorescent dye primer 1 primer 2 primer 3 primer 4
PET UCD-CH11 UCD-CH12 UDP98-411 long BPPCT 034 long
size range (bp) 144–168 176–200 268–284 294–330
NED UCD-CH31 PS12A02 long EMPaS06 PceGA34 long
size range (bp) 124–136 157–179 204–228 239–273
FAM EMPaS11 EMPaS02 EMPaS12 long
size range (bp) 66–108 134–148 224–248
VIC UDP98-412 EMPaS05 UDP97-403 long EMPaS10 long
size range (bp) 113–133 162–174 194–224 281–312
Table 3. Summary evaluation of amplified loci.
locus name k Ne Ho He PIC FNull FIS
EMPaS02 7 3.294 0.542 0.604 0.568 0.0663 0.132
EMPaS10 long 7 3.393 0.614 0.684 0.653 0.0281 0.055
EMPaS11 4 2.678 0.795 0.777 0.737 0.0273 0.069
UCD-CH11 8 2.491 0.771 0.731 0.679 –0.0232 –0.044
UCD-CH31 7 2.898 0.795 0.826 0.799 0.07 0.109
PS12A02 long 10 4.697 0.53 0.592 0.556 0.0201 0.021
UDP98-412 6 4.133 0.723 0.792 0.757 0.0333 0.066
EMPaS05 4 3.643 0.687 0.804 0.778 –0.0027 –0.005
EMPaS06 6 3.371 0.735 0.832 0.805 0.0206 0.023
EMPaS12 long 6 3.401 0.639 0.718 0.671 –0.0933 –0.180
BPPCT 034 long 8 5.789 0.699 0.75 0.702 0.0365 0.068
UDP97-403 long 9 4.820 0.759 0.745 0.703 0.0513 0.106
PceGA34 long 15 4.807 0.744 0.882 0.867 0.0048 0.006
UDP98-411 long 5 3.294 0.602 0.645 0.593 –0.0693 –0.137
UCD-CH12 9 3.934 0.775 0.855 0.834 –0.0101 –0.027
EMPaS02 7 3.294 0.778 0.792 0.76 0.0663 0.132
 
mean 7.400 3.776 0.708 0.729 0.685 0.011 0.017
SD 2.746 0.917 0.081 0.065 0.072 0.046 0.088
SE 0.709 0.237 0.021 0.017 0.019 0.012 0.023
k – number of detected alleles
Ne – effective number of alleles
Ho, He – observed and expected heterozygosity
PIC – polymorphic information content
FNull – potential occurrence of null alleles
FIS – fixation index